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. 2014 Oct 30;10(10):e1004474. doi: 10.1371/journal.ppat.1004474

Figure 1. Abrogation of TRIM5alpha restriction by mixed p12 mutant particles.

Figure 1

(A) Schematic representation of the Gag-Pol expression plasmid and p12 substitution mutants used in this study showing the amino acid sequence of Mo-MLV and N-MLV p12. (B) LacZ-encoding N-MLV VLPs containing either wild type p12, p12 mutant 6 or p12 mutant 14 (triangles), or a mixture of both mutants (circles) were synthesised. The percentage of p12 mutant 14 Gag-Pol expression plasmid in the transfection mix is indicated in brackets for the mixed mutants. Serial dilutions of these VLPs were used to challenge TE671 cells. Four hours later, cells were challenged with a fixed dose of GFP-encoding N-MLV VLPs, and after a further three days, the number of GFP positive cells was measured by flow cytometry. The percentage of GFP positive cells is plotted against LacZ-virus dilution. (C) D17 cells were challenged with equivalent RT-units of the VLPs used in (B). Infectivity was measured by detection of beta-galactosidase activity in a chemiluminescent reporter assay and plotted as a percentage of wild type N-MLV infectivity. These data are representative of multiple independent experiments.